Bio-rad Mouse Typer Isotyping Kit Manual do Utilizador

Mouse Typer®Sub-Isotyping KitInstruction ManualProtocol for Mouse Typer Sub-IsotypingKit (Catalog Number 172-2051) and Mouse Typer Sub Isotyping Panel

Note: When assessing color development using Model 3550,Model 550, or the Model 3550-UV Microplate Reader, washthe bottom of the assay plate thoroughl

Problem Probable Cause Recommended SolutionB. No reaction or weak color development.1. Horseradish a. Improper storage of a. Store peroxidase substra

Problem Probable Cause Recommended SolutionC. Monoclonal 1.Impure sample. 1. Use fresh source of cul-exhibits multiple ture supernatant. Purifysub-is

Section 4References1. Hybridomas, American Type Culture Collection, Rockville, MD(1984).2. Beyer, C. F., J. Immunol. Methods, 67, 79 (1984).3. Langone

Bio-Rad Laboratories, 2000 Alfred Nobel Drive, Hercules, CA 94547LIT78 Rev DLIT78D 8/7/98 04:08 PM Page BC1

Table of ContentsSection 1 Introduction ...11.1 Background...

Section 1Introduction1.1 BackgroundThe growth and widespread use of mouse monoclonalantibody technology have created a need for a fast, accurate, ands

1.2 Materials RequiredKit Components:Catalog Quantity/Number Product Description Package172-2055 Mouse Typer Sub-Isotyping Panel, includes 10 ml each

7. 96-well polystyrene microplates (Bio-Rad catalog number224-0096).8. Pipet tips (Bio-Rad catalog number 223-9302, nonsterile).9. Octapette®pipet, 10

Blocking solution, 1% BSA-PBS.30 ml Add 0.3 g BSA to 30 ml PBS. Adjust pHto 7.2.Antibody conjugate Dilute GAR-HRP conjugate 1:3,000 by solution, 10 m

4. Monoclonal Antibody Sample: To insure proper typing ofculture media samples, do not dilute when applying tomicroplates. When using ascites fluid, d

2. Remove any unbound Ag by flooding the wells of the assayplates with PBS. Use a plastic wash bottle or an automaticplate washer. When using a wash b

Fig. 1. Suggested format for sub-isotyping 10 sampleson 1 microplate.7. Empty the plates of panel reagents and flick-wash 5x withPBS-tween.8. With the